Kirchner Marieluise, Selbach Matthias
Cell Signalling and Mass Spectrometry Group, Max Delbrueck Center for Molecular Medicine, Berlin, Germany.
Methods Mol Biol. 2012;893:175-99. doi: 10.1007/978-1-61779-885-6_13.
Mass spectrometry-based quantitative proteomics can identify and quantify thousands of proteins in complex biological samples. Improved instrumentation, quantification strategies and data analysis tools now enable protein analysis on a genome-wide scale. Particularly, quantification based on stable isotope labeling with amino acids (SILAC) has emerged as a robust, reliable and simple method for accurate large-scale protein quantification. The spectrum of applications ranges from bacteria and eukaryotic cell culture systems to multicellular organisms. Here, we provide a step-by-step protocol on how to plan and perform large-scale quantitative proteome analysis using SILAC, from sample preparation to final data analysis.
基于质谱的定量蛋白质组学能够鉴定和定量复杂生物样品中的数千种蛋白质。如今，改进的仪器、定量策略和数据分析工具使得在全基因组范围内进行蛋白质分析成为可能。特别是，基于氨基酸稳定同位素标记（SILAC）的定量方法已成为一种强大、可靠且简单的大规模蛋白质准确定量方法。其应用范围涵盖从细菌、真核细胞培养系统到多细胞生物体。在此，我们提供一份关于如何使用SILAC计划和进行大规模定量蛋白质组分析的详细方案，内容从样品制备到最终数据分析。
